CN113030037A - Chlorophyll fluorescence kinetic analysis method for male seedlings and female seedlings of torreya grandis under high-temperature stress - Google Patents
Chlorophyll fluorescence kinetic analysis method for male seedlings and female seedlings of torreya grandis under high-temperature stress Download PDFInfo
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Abstract
The invention provides a chlorophyll fluorescence kinetic analysis method for male and female torreya grandis under high-temperature stress, which comprises the following steps: taking original seedlings of 2-year-old torreya trees as stocks, respectively collecting female torreya seedlings and male seedling twigs as scion scions, and taking the scion scions as the research objects after grafting, survival and growth for 2 years; respectively placing the female and male grafted seedlings in an illumination incubator at 25 deg.C, 30 deg.C, 35 deg.C and 40 deg.C, illuminating for 14 h, reducing temperature by 2 deg.C, and placing in dark environment for 10 h; after dark adaptation of the leaves for 30 min, rapid chlorophyll fluorescence induction kinetic curve determination is carried out, 4 plants are respectively determined for male and female grafted seedlings, and each plant is repeatedly determined for 2 times; the data were processed and analyzed and compared using the least significant difference method. The invention reflects the situation that the seedlings can be irreversibly damaged when large-area Chinese torreya grafting seedlings are planted in the production practice process, and the experimental conclusion can further provide theoretical basis for the Chinese torreya grafting seedling planting process.
Description
Technical Field
The invention relates to the field of general botany, in particular to a chlorophyll fluorescence kinetic analysis method for male and female torreya grandis seedlings under high-temperature stress.
Background
Chinese torreya (Torreya grandis cv. 'Merrillii Hu') genus Taxaceae (Taxaceae) genus Torreya (Taxaceae) ((R)Torrey) Evergreen arbor is a kind of vegetative propagation of torreya. After grafting improvement, the method is used for multipurpose excellent economic tree species such as fruits and oil. The minimum annual absolute temperature is-8 to-15 ℃, the average annual temperature is 14.5 to 17.5 ℃, and the consulting literature shows that: under the continuous sunny and hot high-temperature environment in summer,the production of Chinese torreya is seriously influenced, and measures such as tending operation, shading irrigation and the like are not adopted for newly-built Chinese torreya forests to reduce loss. The torreya grandis has sufficient rainwater before blooming, proper temperature, moderate rainwater in the flowering period and the flower falling period, sufficient illumination, sufficient rainwater in the fruit setting period and cool climate in summer, and is favorable for high yield of the torreya grandis. In the high-temperature stress process, the relative membrane permeability, MDA content, SOD, POD, CAT enzyme activity, Pro and SS content of the Chinese torreya leaves take 39 ℃ as peak values, the peak values are gradually reduced after the peak values are continuously increased, when the peak values reach 42 ℃, the Chinese torreya leaves are seriously damaged, and a protective enzyme system is damaged after short-time stress.
The grafting technology is uniformly and widely applied in the production process of the Chinese torreya, and is also one of the measures for improving the yield of the Chinese torreya and enhancing the resistance. In order to comprehensively reflect the situation that the seedlings are irreversibly damaged when large-area Chinese torreya grafting seedlings are planted in the production practice process, the experimental conclusion can further provide theoretical basis for the Chinese torreya grafting seedling planting process, guide production and reduce loss. Moreover, the sensitivity of the grafted female torreya grandis and the grafted male torreya grandis to high temperature in summer is not completely the same, and in order to more accurately guide the cultivation and management of the grafted female torreya grandis and the grafted male torreya grandis, it is necessary to research and analyze the rapid chlorophyll fluorescence induction kinetic parameters and the OJIP curve of the grafted female torreya grandis and the grafted male torreya grandis under high-temperature stress, and discuss the parameters reflected by partial photosystems of the grafted female torreya grandis and the grafted male torreya grandis under high-temperature stress.
Disclosure of Invention
The invention aims to provide a chlorophyll fluorescence kinetic analysis method for male and female Chinese torreya seedlings under high-temperature stress.
In order to achieve the purpose, the invention adopts the following technical scheme:
a chlorophyll fluorescence kinetic analysis method for male and female torreya grandis seedlings under high-temperature stress comprises the following steps:
step 1): taking original seedlings of 2-year-old torreya trees as stocks, respectively collecting female torreya seedlings and male seedling twigs as scion scions, and taking the scion scions as the research objects after grafting, survival and growth for 2 years;
step 2): placing the female and male grafted seedlings in an illumination incubator at 24-26 deg.C, 29-31 deg.C, 34-36 deg.C, and 39-41 deg.C, respectively, controlling the rest variables, and after 13-15 h of illumination, lowering the temperature by 1-3 deg.C, and placing in a dark environment for 9-11 h;
step 3): after dark adaptation of the leaves is carried out for 25-35 min, selecting leaves at the 3 rd to 4 th leaf positions below the top leaves of the new shoots of the plants to carry out rapid chlorophyll fluorescence induction kinetic curve determination, measuring 4 plants of the male and female grafted seedlings respectively, and repeatedly measuring each plant for 2 times, wherein the measured fluorescence parameters are as follows:
FO: initial fluorescence intensity (50 μ s) after dark adaptation; fM: maximum fluorescence intensity (0.3-2 s); fJ: fluorescence intensity when illuminated for 2 ms; fI: fluorescence intensity at 30 ms of illumination;V t≡(Ft-FO)/(FM-FO): relatively variable fluorescence at time t;V J=(FJ-FO)/(FM-FO): relatively variable fluorescence at point J;Ψ O≡ETO/TRO=(1-V J): the trapped exciton transfers electrons into the electron transfer chain beyondQ A -The probability of other electron acceptors of (a);φ Eo≡ETO/ABS=[1-(F O/F M)]Ψ O: quantum yield of absorbed energy for electron transfer;φ Do≡1-φ Po=(F O/F M): quantum ratios for heat dissipation;δ Ro≡REo/ETo=(F M-F I)/(F M-F J): quantum yield of the captured energy transferred to the end of the electron chain; ABS/CSm ≈F M: absorbed light energy per unit area; TRo/CSm =φ Po(ABS/CSm): light energy captured per unit area; ETo/CSm =φ Eo(ABS/CSm): quantum yield of electron transfer per unit area; DIo/CSm = (ABS/CSm) - (TRo/CSm): heat dissipation per unit area; RC/CSm =φ Po(V JMo) (ABS/CSm): the number of reaction centers per unit area; PI (proportional integral)ABS≡(RC/ABS)[φ Po/(1-φ Po)][Ψ O/(1-Ψ O)]: a performance index based on absorbed light energy; PI (proportional integral)total≡PIABS(δ Ro/1-δ Ro): and (4) comprehensive performance indexes.
Step 4): all data were processed and analyzed, and multiple comparisons were performed using the least significant difference method.
Preferably, in the step 2), the female grafted seedlings and the male grafted seedlings are respectively placed in an illumination incubator at the temperature of 25 ℃, 30 ℃, 35 ℃ and 40 ℃, illuminated for 14 hours, then the temperature is reduced by 2 ℃ and the female grafted seedlings and the male grafted seedlings are placed in a dark environment for 10 hours.
Preferably, in step 3), the rapid chlorophyll fluorescence induction kinetic curve is determined using a multifunctional plant efficiency analyzer (M-PEA, Hansatech, Norfolk, UK).
Adopt above-mentioned technical scheme to have following beneficial effect:
the invention researches and analyzes rapid chlorophyll fluorescence induction kinetic parameters and an OJIP curve of a Chinese torreya grafted female seedling and a Chinese torreya grafted male seedling under high-temperature stress, discusses parameters reflected by partial photosystems of the Chinese torreya grafted female seedling and the Chinese torreya grafted male seedling under the high-temperature stress environment, and mainly aims to comprehensively reflect the condition that the Chinese torreya grafted seedlings planted in a large area can cause irreversible damage to seedlings in the production practice process, so that a test conclusion can further provide theoretical basis for the Chinese torreya grafted seedling planting process, guide production and reduce loss.
Drawings
The invention will be further described with reference to the accompanying drawings in which:
FIG. 1 is a comparison of rapid fluorescence induction curves of Torreya grandis grafted male and female seedling leaves stressed by different temperatures.
FIG. 2 shows the leaves F of Torreya grandis grafted with male and female seedlings under different temperature stressesV/FMAnd (5) comparing the values.
FIG. 3 shows the temperature stress of Torreya grandis grafting male and female seedling leaves PIABSAnd (5) comparing the values.
FIG. 4 shows the temperature stress of Torreya grandis grafting male and female seedling leaves PItotalAnd (5) comparing the values.
FIG. 5 is a comparison of net photosynthetic rate values of leaves of Torreya grandis grafted male and female seedlings under different temperature stresses.
FIG. 6 shows the comparison of stomatal conductance values of leaves of male and female torreya grandis grafted by different temperature stresses.
FIG. 7 shows intercellular CO in leaves of Torreya grandis grafted male and female seedlings under different temperature stresses2And (6) comparing concentration values.
FIG. 8 is a comparison of transpiration rate values of leaves of torreya grandis grafted male and female seedlings under different temperature stresses.
Detailed Description
The present invention will be described in further detail with reference to the following drawings and specific examples.
The invention provides a chlorophyll fluorescence kinetic analysis method for male and female torreya grandis seedlings under high-temperature stress, which comprises the following steps:
step 1): taking original seedlings of 2-year-old torreya trees as stocks, respectively collecting female torreya seedlings and male seedling twigs as scion scions, and taking the scion scions as the research objects after grafting, survival and growth for 2 years;
step 2): placing the female and male grafted seedlings in an illumination incubator at 24-26 deg.C, 29-31 deg.C, 34-36 deg.C, and 39-41 deg.C, respectively, controlling the rest variables, and after 13-15 h of illumination, lowering the temperature by 1-3 deg.C, and placing in a dark environment for 9-11 h;
in the embodiment, the female and male grafted seedlings are respectively placed in an illumination incubator at the temperature of 25 ℃, 30 ℃, 35 ℃ and 40 ℃ and are illuminated for 14 hours, and then the temperature is reduced by 2 ℃ and the incubator is placed in a dark environment for 10 hours;
step 3): after dark adaptation of the leaves is carried out for 25-30 min, rapid chlorophyll fluorescence induction kinetic curve determination is carried out, 4 plants are respectively determined for male and female grafted seedlings, each plant is repeatedly determined for 2 times, and the determined fluorescence parameters are as follows: fo: initial fluorescence intensity (50 μ s) after dark adaptation; fM: maximum fluorescence intensity (0.3-2 s); fJ: fluorescence intensity when illuminated for 2 ms; fI: fluorescence intensity at 30 ms of illumination;V t≡(Ft–Fo)/(FM-Fo): relatively variable fluorescence at illumination time t;V J=(FJ–Fo)/(FM-Fo): relatively variable fluorescence at point J;Ψo≡ETo/TRo=(1-V J): the trapped exciton transfers electrons into the electron transfer chain beyondQ A -The probability of other electron acceptors of (a); phi is aEo≡ETo/ABS=[1-(Fo/FM)]Ψ O: quantum yield of absorbed energy for electron transfer;φ Do≡1-φ Po＝(Fo/FM): quantum ratios for heat dissipation;δ Ro≡REo/ETo=(FM-FI)/(FM-FJ): quantum yield of the captured energy transferred to the end of the electron chain; ABS/CSm ≈ FM: absorbed light energy per unit area; TRo/CSm =φ Po(ABS/CSm): light energy captured per unit area; ETo/CSm =φ Eo(ABS/CSm): quantum yield of electron transfer per unit area; DIo/CSm = (ABS/CSm) - (TRo/CSm): heat dissipation per unit area; RC/CSm =φ Po(V JMo) (ABS/CSm): the number of reaction centers per unit area; PI (proportional integral)ABS≡(RC/ABS)[φ Po/(1–φ Po)][Ψ O/(1–Ψ O)]: a performance index based on absorbed light energy; PI (proportional integral)total≡PIABS(δ Ro/1–δ Ro): and (4) comprehensive performance indexes.
In this example, a rapid chlorophyll fluorescence-induced kinetic curve (OJIP) was determined using a multifunctional plant efficiency analyzer (M-PEA, Hansatech, Norfolk, UK). All leaves were subjected to 30 min dark adaptation and subjected to JIP-test analysis on the OJIP curve.
Step 4): all data were processed and analyzed, and multiple comparisons were performed using the least significant difference method.
As shown in figure 1, after high-temperature stress, the rapid chlorophyll fluorescence induction kinetic curve of the Chinese torreya grafted female seedling and male seedling leaves is obviously changed. Within 35 ℃, the initial fluorescence of the two has small change amplitude compared with the normal temperature, and the increase amplitude is large under the high temperature stress of 40 ℃. The J-I phase inflection points of both started to disappear under high temperature stress at 40 ℃. The maximum fluorescence intensity of the two is reduced compared with the normal temperature, the reduction at the high temperature of 40 ℃ is particularly obvious, the J-I phase in the curve almost tends to be smooth but not completely disappears, and the reduction range of the fluorescence output value of the point I and the point P is obvious. The results show that the damage degree of the photosynthetic mechanism of the Chinese torreya grafted female seedling and the male seedling is basically the same under the high-temperature stress.
Fv/Fm reflects an important parameter for the light energy conversion efficiency of the PS II active center. In the figure, different upper/lower case letters indicate that the difference between different varieties is significant (P < 0.05) at different temperatures of the same variety and at the same temperature, and each numerical value is the mean value plus/minus standard deviation, which is the same below. Data from FIG. 2 and Table 1 show F of Torreya grandis graftsV/FMThe value is gradually reduced along with the increasing of the stress degree (P is less than 0.05). Under normal conditions, the treatment at 30 ℃ has no obvious change, and F of male seedlings and female seedlings grafted on Chinese torreya at the temperature of more than 35 DEG CV/FMThe values are obviously reduced compared with the control, and are respectively reduced by 37.3 percent and 25.7 percent; the temperature of 40 ℃ is reduced to the minimum value, the temperature is respectively reduced by 83 percent and 66.7 percent, and the male seedlings are obviously lower than the female seedlings, which shows that electrons flow from the PS II reaction center to the female seedlingsQ A、Q BAnd PQ library transfer is blocked, Fv/Fm value is remarkably reduced, and the electron transfer inhibition in the leaves of the male seedling grafted with the Chinese torreya is higher than that of the female seedling grafted with the Chinese torreya.
PIABSIs a performance index based on the absorption of light energy. As seen from FIG. 3 and Table 1, the photosynthetic performance index of the grafted torreya grandis seedlings is remarkably reduced under the high-temperature stress (40 ℃) environment. From the beginning of the environment culture at 25 ℃, the photosynthetic performance indexes of the two show the trend of decreasing after increasing, the highest value is reached at about 30 ℃, the performance index of the male seedling grafted by the Chinese torreya is always lower than that of the female seedling, and the difference is obvious. Under the high temperature stress of 40 ℃, the photosynthetic performance indexes of the two are obviously reduced, and the PI is reflectedABSSusceptibility to high temperature stress.
Comprehensive performance index PItotalMainly researches the influence of high-temperature stress on the leaves PS I of the torreya grandis grafted seedlings by the electron transfer activity of the torreya grandis grafted seedling optical system. As can be seen from FIG. 8 and Table 1, under the stress of high temperature environment, the Chinese torreya grafts the female and male seedlings PItotalAnd is significantly reduced. The two are cultured in an environment of 25 ℃, the comprehensive performance index shows a trend of decreasing after increasing, and the maximum value is reached at about 30 ℃. The comprehensive performance index of the male grafting Chinese torreya seedlings is always higher than that of the female grafting Chinese torreya seedlings, and the difference is obvious.
TABLE 1 analysis of difference among maximum photochemical efficiency, photosynthetic performance index and comprehensive performance index of leaves of Torreya grandis grafted male and female seedlings under high temperature stress
As can be seen from Table 2, at normal temperature, the unit area of the grafted female torreya grandis seedlings can absorb higher light energy than the unit area of the grafted male torreya grandis seedlings, the absorbed light energy is more, and the difference is obvious. Under the stress of high temperature of 30 ℃, 35 ℃ and 40 ℃, the light energy absorbed (ABS/CSm), the captured light energy (TRo/CSm) and the transmitted light energy (ETo/CSm) in a unit area all rise first and then fall, and the parts of electrons absorbed, captured and transmitted by the grafted female seedling are obviously higher than those of the grafted male seedling. The heat dissipation per unit area (DIo/CSm) is obviously increased gradually under the environment of gradually increasing temperature, and the data in the table 2 show that the energy dissipation of the grafted female seedlings is lower than that of the grafted male seedlings. The fact that the energy share of the torreya grandis leaves for electron transfer is reduced, the electron transfer is blocked, the energy share of heat dissipation of the torreya grandis leaves and the torreya grandis leaves is increased, and the electron blocking degree of the grafted female seedling leaves is lower than that of the grafted male seedling leaves. In general, the ratio of the quantum efficiency parameter per unit area of the Chinese torreya grafted female seedling and the Chinese torreya grafted male seedling to the control temperature under high temperature stress has large change and obvious difference, wherein the change of the energy flow parameter per unit area of the Chinese torreya grafted female seedling is smaller than that of the Chinese torreya grafted male seedling after the high temperature stress.
TABLE 2 difference between energy flow parameters per cross-sectional area and reaction center density of Torreya grandis grafted female and male seedlings under high temperature stress
V JReflecting the degree of closing of the active reaction center of the photosystem II, and grafting the Chinese torreya female seedlings and male seedlings at the high temperature of 40 ℃ under the stress of the normal temperatureV JRespectively increased by 123.76% and 324.55%.
Grafting female seedling and male seedling leaves on torreya grandis after high-temperature stressΨo andφeo gradually decreases, and the difference between the decreasing amplitude and the normal temperature is obvious. Grafted female seedling leafSheetΨo andφthe decrease in Eo was less gradual than that of the male seedlings, but the difference was not significant. The heat dissipation quantum ratio of the grafted female seedling and the grafted male seedling is respectively increased by 257.95% and 279.09%, and the increase value of the female seedling is lower than that of the male seedling, but the difference is not obvious.
TABLE 3 difference in Vj and quantum yield under high temperature stress of Torreya grandis grafted female and male seedling leaves
Measuring gas exchange parameters of torreya grandis leaves by using a portable photosynthetic system, wherein the gas flow during measurement is 500 mu mol s-1Under the measurement condition of CO2The concentration is 400 +/-5 mu mol-1Measuring net photosynthetic rate (Pn), stomatal conductance (Gs) and intercellular CO of Chinese torreya grafted female seedling and male seedling leaves2Concentration (Ci), transpiration rate (Tr) and other parameters, and the determination time is 11: 00-13: 00 at noon of 3 days under continuous stress at different temperatures.
TABLE 4 difference in gas exchange parameters between Torreya grandis grafted female and male seedlings under high temperature stress
The data and comparative plots of table 4 and fig. 5-8 show: under the stress of different temperatures, net photosynthetic rate values of leaves of the Chinese torreya grafted female seedlings and male seedlings are increased and then decreased, the net photosynthetic rate of the female seedlings is higher than that of the male seedlings, the net photosynthetic rate of the female seedlings and the male seedlings reach the highest values at about 30 ℃, and the difference is obvious from 35 ℃. The stomatal conductance value is continuously and obviously reduced from 25 ℃, and the stomatal conductance of female seedlings is slightly higher than that of male seedlings. Intercellular CO2The concentration is higher than 30-35 ℃ in room temperature environment, and intercellular CO is in high temperature environment2The concentration is obviously increased, and intercellular CO of male and female grafted seedling leaves at various temperatures2The concentration difference is significant. The transpiration rates of the two leaves at different temperatures are gradually reduced along with the temperature rise, and the transpiration rates of the female seedling leaves are obviously reduced, and are slightly higher than those of male seedlings. The gas exchange parameters of the test subject blades show that:the gas exchange capacity of the leaves of the Chinese torreya grafted female seedling in a high-temperature environment is stronger than that of the grafted male seedling.
PS II is the region of the plant photosynthetic system most susceptible to high temperature stress. The PS II inhibition condition of the torreya grandis seedlings under continuous high-temperature treatment is researched, and the adaptability of the torreya grandis seedling leaves to a high-temperature environment can be reflected. The change of plant thylakoid membrane can be quickly captured by chlorophyll fluorescence, and the main reasons of K point of chlorophyll fluorescence quick induction curve are the cracking process of water and part of electron primary quinone acceptor (B)Q A) Inhibited, indicating that the oxygen-releasing complex is somewhat damaged. In the research, the OJIP curves of the female and male torreya grandis grafted under high temperature stress are greatly changed compared with the room temperature, the maximum fluorescence intensity (P) of the female seedling is higher than that of the male seedling, the O-J-I curves of the female seedling and the male seedling tend to be horizontal straight lines under the environmental stress of 40 ℃, the indexes of the female seedling and the male seedling reach the maximum values at 35 ℃, the temperature gradually rises and then falls, and the temperature falls obviously under the high temperature stress of 40 ℃. The initial fluorescence of the female seedling is obviously higher than that of the male seedling under the stress of high temperature of 40 ℃, the curve is more gentle compared with that of the male seedling, and the J-I phase inflection point of the female seedling disappears later than that of the male seedling. The decrease range of the I and P point fluorescence yield of the female seedling under various temperature environments is smaller than that of the male seedling, and the decrease range is more obvious particularly at 40 ℃. Further indicates that the damage of high temperature stress to the Chinese torreya grafting female seedling is less than that of the male seedling.
The result shows that the damage degree of the photosynthetic mechanism of the Chinese torreya grafted female seedling is lower than that of the grafted male seedling under the high-temperature stress. The OJIP curve can visually reflect the adverse damage degree of plants, and the high temperature stress resistance of the Chinese torreya grafted female seedling is stronger than that of the male seedling.
In this experiment, the high temperature not only destroys the electron transfer on the donor side of the leaf, but also suppresses and reduces the electron transfer rate on the acceptor side of the PS II. The experimental result is analyzed by JIP-test, after the Chinese torreya grafted female seedling and male seedling are stressed by high temperature,V Jthe rise was significant indicating that electron transport was affected. Both are stressed at a high temperature of 40 DEG CΨo is significantly reduced, resulting inφEo drops, the test subject consumes too many of the activated electrons,φthe Do is significantly elevated. From the test data, the temperature is increased by 40 DEG CAfter stress, female seedling is grafted on Chinese torreyaV JThe ascending amplitude is small, the energy consumption is low, more electrons enter a transfer chain, the transfer capability of a receptor electron in a leaf photosystem II is higher than that of a Chinese torreya grafting male seedling, and the capability of a photosynthetic mechanism of the system to cope with a high-temperature stress environment is higher than that of a grafting male seedling.
In the research, the temperature stress causes the Fv/Fm reduction of the Chinese torreya grafted female seedlings and male seedlings, which accords with the research result of plants subjected to high-temperature stress, and shows that the efficiency of electron transfer is reduced by converting the light energy of PS II after the high-temperature stress. The Fv/Fm value of the Chinese torreya grafted female seedling is reduced less than that of the male seedling after high-temperature stress, which indicates that the light energy conversion rate and the electron transfer efficiency of the Chinese torreya grafted female seedling PS II in a high-temperature environment are higher than those of the grafted male seedling under the same condition. Under the stress of temperature, the distribution and the change of energy flow of reaction centers of the two leaves are obvious, and the reduction range of the Torreya grandis grafting female seedlings ABS/CSm, TRo/CSm and ETo/CSm is obviously higher than that of the grafting male seedlings.
PIABSMainly reflecting the efficiency of the PS II reaction center. Research suggests that PIABSThe running state of a plant photosynthetic mechanism can be accurately reflected, and the plant photosynthetic mechanism is sensitive to adversity stress ratio Fv/Fm. In this study, Torreya grandis grafted female and male seedling leaves PIABSFv/Fm is sensitive to temperature changes, but the photosynthetic performance index is much sensitive to this. Meanwhile, PIabs and PItotal appear at the same time, so that the problem that the optical system I cannot be reflected is solved, and the transmission capability between the optical system I and the optical system II and other related performances are reflected. The high temperature stress of the time is different from the conditions of rapid determination after 2h of high temperature stress on the plants in the past, and the like, the plants are subjected to simulation experiment by adopting a continuous high temperature stress environment, and PI is performed at the temperature of 30 ℃ and the normal temperatureABSAnd PItotalAll are increased, PIABSThe increase is significant and both show a tendency to decrease gradually with increasing temperature, which is mainly related to the gradual destruction of the photosynthetic machinery, which is consistent with the trend of Fv/Fm. From the data, the Torreya grandis grafted female seedling is more PI than the grafted male seedling under the stress of high-temperature environmentABSAnd PItotalThe above is good.
In the experimental process, the temperature change causes the gas of the leaves of the male and female seedling grafted by the Chinese torreyaThe change of the exchange parameters, along with the rise of the temperature, the net photosynthetic rate (Pn), the stomatal conductance (Gs) and the transpiration rate (Tr) are gradually reduced to accord with the research result of the plants after high-temperature stress, which shows that the photosynthesis of the plants is inhibited. Intercellular CO2The concentration (Ci) is damaged due to the photosynthetic mechanism of torreya grandis leaves along with the rise of temperature, the respiration is continuous, the photosynthesis is weakened, and intercellular CO is caused2The concentration increases. The experimental data show that: under the room temperature environment, the Pn, Gs and Tr values of the torreya grandis grafted male and female seedling leaves are not remarkably different and are kept at a certain level. In the process of increasing the temperature, except that Pn slightly increases at about 30 ℃, all numerical values are obviously reduced in a high-temperature environment, and the three numerical values of the Chinese torreya grafted female seedling are higher than those of the Chinese torreya grafted male seedling in each temperature environment. Intercellular CO between them2The concentration is lowest in the range of 30-35 ℃, which shows that the difference of respiration action of photosynthesis is closest in the temperature range, and intercellular CO of the Chinese torreya grafted female seedling is the closest in the whole temperature change process2The concentration is obviously lower than that of the grafted male seedlings.
In conclusion, the photosynthetic performance of the grafted Chinese torreya seedlings is the most stable in the environment of about 30 ℃; through experimental data and discussion and a rapid chlorophyll fluorescence induction kinetic curve, the adaptability of the Chinese torreya grafted female seedling to high temperature is stronger than that of the grafted male seedling, and the application range of the female grafted seedling is wider than that of the male grafted seedling in the production practice process.
Other embodiments of the present invention than the preferred embodiments described above will be apparent to those skilled in the art from the present invention, and various changes and modifications can be made therein without departing from the spirit of the present invention as defined in the appended claims.
Claims (3)
1. A chlorophyll fluorescence kinetic analysis method for male and female torreya grandis seedlings under high-temperature stress is characterized by comprising the following steps of:
step 1): taking original seedlings of 2-year-old torreya trees as stocks, respectively collecting female torreya seedlings and male seedling twigs as scion scions, and taking the scion scions as the research objects after grafting, survival and growth for 2 years;
step 2): placing the female and male grafted seedlings in an illumination incubator at 24-26 deg.C, 29-31 deg.C, 34-36 deg.C, and 39-41 deg.C, respectively, controlling the rest variables, and after 13-15 h of illumination, lowering the temperature by 1-3 deg.C, and placing in a dark environment for 9-11 h;
step 3): after dark adaptation of the leaves is carried out for 25-35 min, selecting leaves at the 3 rd to 4 th leaf positions below the top leaves of the new shoots of the plants to carry out rapid chlorophyll fluorescence induction kinetic curve determination, measuring 4 plants of the male and female grafted seedlings respectively, and repeatedly measuring each plant for 2 times, wherein the measured fluorescence parameters are as follows:
FO: initial fluorescence intensity after dark adaptation; fM: the maximum fluorescence intensity; fJ: fluorescence intensity when illuminated for 2 ms; fI: fluorescence intensity at 30 ms of illumination;V t≡(Ft-FO)/(FM-Fo): relatively variable fluorescence at time t;V J=(FJ-FO)/(FM-FO): relatively variable fluorescence at point J;Ψ O≡ETO/TRO=(1-V J): the trapped exciton transfers electrons into the electron transfer chain beyondQ A -The probability of other electron acceptors of (a);φ Eo≡ETO/ABS=[1-(FO/FM)]Ψ O: quantum yield of absorbed energy for electron transfer;φ Do≡1-φ Po=(FO/FM): quantum ratios for heat dissipation;δ Ro≡REo/ETo=(FM-FI)/(FM-FJ): quantum yield of the captured energy transferred to the end of the electron chain; ABS/CSm ≈F M: absorbed light energy per unit area; TRo/CSm =φ Po(ABS/CSm): light energy captured per unit area; ETo/CSm =φ Eo(ABS/CSm): quantum yield of electron transfer per unit area; DIo/CSm = (ABS/CSm) - (TRo/CSm): heat dissipation per unit area; RC/CSm =φ Po(V JMo) (ABS/CSm): the number of reaction centers per unit area; PI (proportional integral)ABS≡(RC/ABS)[φ Po/(1-φ Po)][Ψ O/(1-Ψ O)]: a performance index based on absorbed light energy; PI (proportional integral)total≡PIABS(δ Ro/1-δ Ro): a comprehensive performance index;
step 4): all data were processed and analyzed, and multiple comparisons were performed using the least significant difference method.
2. The fluorescence kinetic analysis method for male and female torreya grandis seedlings under high-temperature stress according to claim 1, wherein in step 2), the grafted female seedlings and the grafted male seedlings are respectively placed in an illumination incubator at 25 ℃, 30 ℃, 35 ℃ and 40 ℃, after 14 hours of illumination, the temperature is reduced by 2 ℃ and the incubator is placed in a dark environment for 10 hours.
3. The method for analyzing chlorophyll fluorescence kinetics of male and female torreya grandis under high-temperature stress according to claim 1, wherein in step 3), a rapid chlorophyll fluorescence induction kinetics curve is determined by using a multifunctional plant efficiency analyzer.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113607909A (en) * | 2021-08-09 | 2021-11-05 | 中国科学院合肥物质科学研究院 | Method for constructing water body comprehensive toxicity characterization parameters by algae photosynthetic inhibition method |
| CN115839937A (en) * | 2022-12-20 | 2023-03-24 | 江苏省中国科学院植物研究所 | Taxus chinensis stress detection method based on chlorophyll fluorescence imaging technology |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2006132691A (en) * | 2006-09-13 | 2008-03-20 | ООО"Генна и клеточна терапи " (RU) | METHOD FOR FLUOROMETRIC DETERMINATION OF PHOTOSYNTHESIS PHOTOSYNTOTROPHIC ORGANISMS PARAMETERS, DEVICE FOR ITS IMPLEMENTATION AND MEASUREMENT CAMERA |
| CN104099023A (en) * | 2014-07-04 | 2014-10-15 | 中国林业科学研究院林产化学工业研究所 | Pimaric acid type resin acid product and method for extracting pimaric acid type resin acid product from torreya grandis aril |
| CN110907413A (en) * | 2019-10-31 | 2020-03-24 | 南京农业大学 | Method for identifying salt tolerance of cabbage mustard by utilizing chlorophyll fluorescence technology |
| CN111802155A (en) * | 2020-07-14 | 2020-10-23 | 浙江农林大学暨阳学院 | Azalea high-temperature resistance evaluation method based on chlorophyll fluorescence OJIP curve |
-
2021
- 2021-02-07 CN CN202110174626.4A patent/CN113030037B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2006132691A (en) * | 2006-09-13 | 2008-03-20 | ООО"Генна и клеточна терапи " (RU) | METHOD FOR FLUOROMETRIC DETERMINATION OF PHOTOSYNTHESIS PHOTOSYNTOTROPHIC ORGANISMS PARAMETERS, DEVICE FOR ITS IMPLEMENTATION AND MEASUREMENT CAMERA |
| CN104099023A (en) * | 2014-07-04 | 2014-10-15 | 中国林业科学研究院林产化学工业研究所 | Pimaric acid type resin acid product and method for extracting pimaric acid type resin acid product from torreya grandis aril |
| CN110907413A (en) * | 2019-10-31 | 2020-03-24 | 南京农业大学 | Method for identifying salt tolerance of cabbage mustard by utilizing chlorophyll fluorescence technology |
| CN111802155A (en) * | 2020-07-14 | 2020-10-23 | 浙江农林大学暨阳学院 | Azalea high-temperature resistance evaluation method based on chlorophyll fluorescence OJIP curve |
Non-Patent Citations (3)
| Title |
|---|
| CAKMAK I. ET AL.: "Magnesium deficiency and high light intensity enhance activities of superoxide dismutase, ascorbate peroxidase, and glutathione reductase in bean leaves", PLANT PHYSIOL. * |
| 刘倩倩;马寿宾;冯希环;孙艳;衣艳君;刘维信;: "嫁接对高温和低温胁迫下辣椒幼苗快速叶绿素荧光诱导动力学特性的影响", 园艺学报, no. 05 * |
| 季永亮 等: "高温胁迫对香榧和榧树幼苗抗氧化系统的影响", 分子植物育种, vol. 18, no. 24 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113607909A (en) * | 2021-08-09 | 2021-11-05 | 中国科学院合肥物质科学研究院 | Method for constructing water body comprehensive toxicity characterization parameters by algae photosynthetic inhibition method |
| CN113607909B (en) * | 2021-08-09 | 2023-11-07 | 中国科学院合肥物质科学研究院 | Construction method of comprehensive toxicity characterization parameters of water body by algae photosynthesis inhibition method |
| CN115839937A (en) * | 2022-12-20 | 2023-03-24 | 江苏省中国科学院植物研究所 | Taxus chinensis stress detection method based on chlorophyll fluorescence imaging technology |
| CN115839937B (en) * | 2022-12-20 | 2024-01-09 | 江苏省中国科学院植物研究所 | Taxus chinensis stress detection method based on chlorophyll fluorescence imaging technology |
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